Optical Activation of TrkA Signaling
Optical Activation of TrkA Signaling
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DOI：
10.1021/acssynbio.8b00126
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发表时间：
2018-07-01
影响因子：
4.7
通讯作者：
Cui, Bianxiao
中科院分区：
文献类型：
--
作者：
Duan, Liting;Hope, Jen M.;Cui, Bianxiao
神经生长因子/原肌球蛋白受体激酶A（NGF/TrkA）信号传导在神经元发育、功能、存活和生长中起关键作用。该通路与神经退行性疾病有关，包括阿尔茨海默病、慢性疼痛、炎症和癌症。NGF结合TrkA的细胞外结构域，导致受体细胞内激酶结构域的激活。由于TrkA信号是高度动态的，机制研究将受益于具有高空间和时间分辨率的工具。在这里，我们提出了四个策略的设计和评估光诱导激活的TrkA在没有神经生长因子。我们的策略涉及光敏感蛋白拟南芥隐花色素2及其结合伙伴CIB 1。我们证明了成功的重演天然NGF/TrkA功能的质膜招聘和TrkA的细胞内结构域的同源相互作用的光诱导。这种方法激活PI 3 K/AKT和Raf/ERK信号通路，促进PC 12细胞中的神经突生长，并在缺乏NGF的情况下支持背根神经节神经元的存活。这种使用光激活TrkA的能力赋予了研究NGF/TrkA信号传导的高空间和时间分辨率。
Nerve growth factor/tropomyosin receptor kinase A (NGF/TrkA) signaling plays a key role in neuronal development, function, survival, and growth. The pathway is implicated in neurodegenerative disorders including Alzheimer's disease, chronic pain, inflammation, and cancer. NGF binds the extracellular domain of TrkA, leading to the activation of the receptor's intracellular kinase domain. As TrkA signaling is highly dynamic, mechanistic studies would benefit from a tool with high spatial and temporal resolution. Here we present the design and evaluation of four strategies for light-inducible activation of TrkA in the absence of NGF. Our strategies involve the light-sensitive protein Arabidopsis cryptochrome 2 and its binding partner CIB1. We demonstrate successful recapitulation of native NGF/TrkA functions by optical induction of plasma membrane recruitment and homo-interaction of the intracellular domain of TrkA. This approach activates PI3K/AKT and Raf/ERK signaling pathways, promotes neurite growth in PC12 cells, and supports survival of dorsal root ganglion neurons in the absence of NGF. This ability to activate TrkA using light bestows high spatial and temporal resolution for investigating NGF/TrkA signaling.